CFSE staining problem with PBMC

From: Jeffrey Dorfman (jdorfman@kilifi.mimcom.net)
Date: Wed Oct 09 2002 - 04:31:39 EST


Dear All,
I have been having a problem staining PBMC with CFSE for cell division
tracking.

Whenever I label PBMC, I seem to get a second population that is dimmer than
the rest of the cells. They do not seem to be cells that have divided, as I
see this even after an overnight incubation with no overt stimulation.

The cells are not T cells and have a FSC/90LS profile matching that of
lymphocytes. Often, these dim cells are as much as 20% of undivided cells in
the lymphocyte FSC/90LS gate and they persist without apparent division for
as long as I have tested: one week.

I have seen this with two batches of CFSE, with a range of concentrations
and labelling times of CFSE and with PBMC from a variety of donors.


Is anyone familiar with what causes this/what these cells are and more
importantly, what conditions to use to prevent this? I have never seen such
a thing when labelling a large variety of mouse cell types with CFSE.

Thank you for any help you can render.

Jeff Dorfman


------
Jeffrey Dorfman, Ph.D.
Kenya Medical Research Institute
Wellcome Trust Research Laboratories
Post Office Box 230
Kilifi, Kenya

jdorfman@kilifi.mimcom.net
jeffreydorfman@yahoo.com

Phone +254 125 22063
Fax   +254 125 22390
Mobile+254 733 86.23.10
Home  +254 125 22053

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